文章摘要
王 静,张国新.OPN通过SDF-1/CXCR4轴调控胃癌细胞的生物学行为[J].南京医科大学学报,2013,(8):1055~1059
OPN通过SDF-1/CXCR4轴调控胃癌细胞的生物学行为
Osteopontin regulates the biological behaviour of gastric cancer cell via SDF-1/CXCR4 axis
投稿时间:2012-10-08  
DOI:10.7655/NYDXBNS20130806
中文关键词: OPN  CXCR4  MMP2  胃癌  迁移
英文关键词: OPN  CXCR4  MMP2  gastric cancer  migration
基金项目:国家自然科学基金(30770992)
作者单位
王 静 苏州市吴中人民医院消化科,江苏 苏州 215000 
张国新 南京医科大学第一附属医院消化科,江苏 南京 210029 
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中文摘要:
      目的:观察骨桥蛋白(osteopontin,OPN)小干扰RNA稳定转染人胃癌细胞SGC-7901后,细胞增殖?凋亡及迁移能力的变化并探讨其可能的分子机制。方法:用脂质体法分别将OPNsiRNA-pcDNATM 6.2及空载体质粒pcDNATM6.2转染人胃癌细胞SGC-7901,稻瘟素筛选,克隆环挑取细胞克隆,用Western blot及RT-PCR技术筛选阳性克隆,并检测稳转细胞中OPN?趋化因子受体4(CXCR4)?基质金属蛋白酶2(MMP2)的表达,应用MTT法?流式细胞仪?细胞迁移试验分别检测转染细胞的增殖?凋亡及迁移能力。结果:OPN siRNA稳定转染细胞SGC-7901后,稳转细胞中CXCR4和MMP2表达量下降;细胞的增殖能力(与转染空载体质粒比较)降低?细胞凋亡率显著高于对照组,转染细胞的迁移能力明显降低。结论:OPN siRNA使胃癌细胞增殖和迁移能力降低?细胞凋亡增加;提示OPN可能通过SDF-1/CXCR4轴及MMP2参与的信号通路调节肿瘤细胞生长及转移。
英文摘要:
      Objective:To observe the biological changes of human gastric cancer cell line SGC-7901, which was stably transfected with siRNA targeting osteopontin (OPN) and to study the molecular mechanism related to the biological changes. Methods:SGC-7901 cells were transfected with OPNsiRNA-pcDNATM6.2 and pcDNATM6.2 by lipofectamine 2000. Transfectants were selected and confirmed by Western blot and RT-PCR technique. The proliferation activity was detected by MTT assay and cell apoptosis was tested by flow cytometry. The migration of transfected cells was assayed by using transwell migration chambers. The expressions of OPN,CXCR4,MMP2 on mRNA and protein level were determined by RT-PCR and Western blot. Result:SGC-7901 cells were stably transfected with OPNsiRNA-pcDNATM6.2. MTT showed that the growth of OPNsiRNA transfected cells was slower than that of empty vecter transfected cells. OPNsiRNA gene transfection can increase the apoptosis. The migration of cells transfected with OPNsiRNA was suppressed compared with the cells transfected with empty vector,and the expressions of CXCR4 and MMP2 in SGC-7901 cells were significantly suppressed after OPN silenced by RNA interference. Conclusion:After OPN siRNA treatment,the cell proliferation and motility are suppressed,and apoptosis is enhanced;CXCR4 and MMP2 are OPN targeting genes. OPN may regulate the growth and metastasis of tumor cell via SDF-1/CXCR4 axis and MMP2 involved signaling pathway.
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